Fermentation of low-salt miso as affected by supplementation with ethanol

Steam-cooked soybeans and rice koji were combined (1:1, w/w), mixed with 5% (w/w) NaCl and ground into a fine paste. Samples (30 g) were deposited in nylon/polyethylene plastic bags and supplemented with 10 ml of aqueous ethanol solutions to give concentrations of 0, 2.5, 5, 7.5, 10, 15, 20, and 25% ethanol. Mixtures were homogenized, sealed, and incubated at 28°C for eight weeks. Mold populations were less than 3 log10 CFU/g in all miso products after four weeks of fermentation. Yeast populations increased to 6.1 log10 CFU/g in the control (0% added ethanol) during the first week of fermentation and remained stable throughout the eight-week fermentation period. Yeasts were not detected in products containing 5–25% ethanol. Populations of lactic acid bacteria (LAB) increased to 6 log10 CFU/g after one week of fermentation in products containing 0 and 2.5% ethanol. However, after eight weeks of fermentation, LAB populations in all products were less than 4 log10 CFU/g. Rapid decreases in pH occurred only in products supplemented with 0 or 2.5% ethanol. Percentages of soluble protein in miso products containing various ethanol concentrations during the eight-week fermentation period revealed that protease activity was still active or not greatly inhibited in products supplemented with less than 10% ethanol. In comparison, koji enzymes were comparatively less affected by ethanol than were populations of molds, yeasts, and LAB. Total soluble carbohydrate and glucose contents were higher in products supplemented with 5, 7.5 and 10% ethanol than in other products. Discoloration (browning) during fermentation occurred most rapidly in products supplemented with 5 or 7.5% ethanol. Sensory evaluation of the low-salt (5%) product supplemented with 7.5% ethanol and fermented for eight weeks revealed normal or enhanced flavor ratings compared to ratings for a commercial product.